US8092608B2 - Composition comprising a prebiotic for decreasing inflammatory process and abnormal activation of non-specific immune parameters - Google Patents

Composition comprising a prebiotic for decreasing inflammatory process and abnormal activation of non-specific immune parameters Download PDF

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US8092608B2
US8092608B2 US10/472,731 US47273104A US8092608B2 US 8092608 B2 US8092608 B2 US 8092608B2 US 47273104 A US47273104 A US 47273104A US 8092608 B2 US8092608 B2 US 8092608B2
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prebiotic
fos
composition
elderly
specific immune
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Florence Rochat
Eduardo Schiffrin
Yves Guigoz
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Nestec SA
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/70—Carbohydrates; Sugars; Derivatives thereof
    • A61K31/702—Oligosaccharides, i.e. having three to five saccharide radicals attached to each other by glycosidic linkages
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
    • A23K50/42—Dry feed
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K50/00—Feeding-stuffs specially adapted for particular animals
    • A23K50/40—Feeding-stuffs specially adapted for particular animals for carnivorous animals, e.g. cats or dogs
    • A23K50/48—Moist feed
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/40—Complete food formulations for specific consumer groups or specific purposes, e.g. infant formula
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/66—Microorganisms or materials therefrom
    • A61K35/74—Bacteria
    • A61K35/741—Probiotics
    • A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
    • A61K35/745—Bifidobacteria
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/04—Drugs for disorders of the alimentary tract or the digestive system for ulcers, gastritis or reflux esophagitis, e.g. antacids, inhibitors of acid secretion, mucosal protectants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs

Definitions

  • the present invention relates to a composition comprising prebiotic (prebiotic adjuvant) for decreasing inflammatory process by improving the homeostasis of non-specific immune defense parameters and of lymphocyte subpopulations. It also relates to the use of a prebiotic formulation in the manufature of a medicament or food or petfood composition for decreasing inflammatory process and/or abnormal activation of non-specific immune parameters, such as phagocytes.
  • prebiotics comprise carbohydrates and more specifically, oligosaccharides. Furthermore it is known that they have widely been used as functional food ingredients. They resist hydrolysis by enzymes of the human digestive tract, can reach the colon undegraded and provide a carbohydrate substance particulary suited to the growth of bifidobacteria
  • Oligosaccharides may be produced from glucose, galactose, xylose, maltose, sucrose, lactose, starch, xylan, hemicellulose, inulin, gum or a mixture thereof. Purified commercially available products such as fructooligosaccharides contain greater than about 95% solids in the form of oligosaccharides.
  • Fructooligosaccharides have been studied in humans mainly for functional claims related to the bioavailability of minerals, lipid metabolism and and regulation of bowel habits (Roberfroid, M. B. Delzenne, N. M. Annu Rev Nutr 1998;18:117-143). Little attention has been given to their effect on immunological functions, while indications for modifications of carcinogenesis and stimulation of gut-associated lymphoid tissue comes from animal studies (Pierre, F., et al. Cancer Res 1997;57:225-228) .
  • fructooligosaccharides long (inulin) and short chain (oligofructose) are among the carbohydrates, which escape digestion in the upper gastro-intestinal tract. They are then fermented in the colon and selectively stimulate the growth of bifidobacteria.
  • Human intestinal flora with its important metabolic activity is possibly associated with many health related functions such as maintenance of gut homeostasis, metabolism of xenobiotics and stimulation of gut immunity. It is influenced by disease, diet, stress and possibly ageing.
  • the large intestine contains up to 10 12 bacteria/g faeces with about 10 3 different species from approximately 40-50 genera of bacteria. Most of them are obligate anaerobes with a large population, however, of facultative anaerobes.
  • the main anaerobe species are Bactero ⁇ des , bifidobacteria, eubacteria, which make up to 99% of the total faecal flora, followed by clostridia, lactobacilli and gram positive cocci, enterococci, coliforms, methanogens and at much lower levels sulfate-reducing bacteria (Hill, M. J. Normal gut bacterial flora. 1995;3-17).
  • Gut flora can be divided into species that have beneficial effects, such as bifidobacteria, or harmful effects, such as Pseudonioinas aeruginosa, Protezus species, staphylococci, some clostridia and Veilonellae , and species that are intermediate in effect such as enterococci, Escherichia coli , Enterococci and Bactero ⁇ des .
  • Bifidobacteria and lactobacilli have been reported to have beneficial effects on specific immune functions (Schiffrin, E. J., et al. J Dairy Sci 1995;78:491-497).
  • the present invention aims to provide another composition able to limit the dysregulation of the immune function, and more particularly the abnormal activation of non-specific immune response such as the phagocytes and the monocyte macrophage cell system as well as preserve lymphocyte subpopulations in normal level of activation.
  • the present invention provides a composition comprising at least one prebiotic for decreasing inflammatory process and/or abnormal activation of non-specific immune parameters.
  • It may be particularly intended for decreasing abnormal activation of phagocytes, for example.
  • a prebiotic supplementation can induce decrease in inflammatory process, and particularly can induce changes in non-specific immunity, such as decreased phagocytic activity, as well as a decreased expression of interleukin-6 mRNA in peripheral blood monocytes.
  • the invention provides the use of at least one prebiotic in the manufature of a medicament or a food or pet food composition for decreasing inflammatory process in a mammal.
  • the invention provides use of at least one prebiotic in the manufature of a medicament or a food or pet food composition for decreasing abnormal activation of non-specific immune parameters in a mammal.
  • the invention provides a method of decreasing inflammatory process in a mammal, which comprises administering an effective amount of a prebiotic or composition comprising at least one prebiotic.
  • the invention provides a method of decreasing abnormal activation of non-specific immune parameters in a mammal, which comprises administering an effective amount of a prebiotic or composition comprising at least one prebiotic.
  • An advantage of the present invention is that it provides a decrease in inflammatory process, particularly a decrease expression of Interleukine-6 mRNA in peripheral blood mononuclear cells.
  • Another advantage of the present invention is that it provides a decrease in phagocytic activity of granulocytes and monocytes, particularly in frail patient with chronic inflammatory situation.
  • Yet another advantage of the present invention is that it may be used to improve the inflammatory situation in a mammal and thus reduce the risk of development of deleterious infections, by simple consumption of a food composition according to the present invention. It will be appreciated that intravenous or subcutaneous administration of a drug requires expertise, and compared to oral administration it is not as safe, convenient or acceptable to the patient. In the light of these concerns, the invention provides the clear advantage of a nutritional and/or therapeutic product which may be administered orally.
  • the composition preferably comprises at least one prebiotic or a prebiotic mixture.
  • the prebiotic comprises an oligosachharide produced from glucose, galactose, xylose, maltose, sucrose, lactose, starch, xylan, hemicellulose, inulin, gum (acacia gum, for example) or a mixture thereof.
  • the oligosaccharide comprises fructooligosaccharide (FOS).
  • the prebiotic comprises a mixture of fructooligosaccharide and inulin.
  • this mixture comprises PREBIO1® or a mixture of commercially available RAFTILOSE® and RAFTILINE®.
  • the prebiotic comprises about 50% to about 95% FOS. More preferably it comprises about 60% to about 80% FOS. Most preferably it comprises about 70% FOS.
  • the prebiotic comprises about 10% to about 50% inulin. More preferably it comprises about 20% to about 40% inulin. Most preferably it comprises about 30% inulin.
  • the prebiotic may comprise a mixture of fructooligosaccharides and inulin in the amounts by weight of 70% fructooligosaccharides and 30% inulin.
  • the composition comprises a probiotic in addition to the prebiotic.
  • the probiotic may be Bifidobacterium bifidum or Streptococcus thermophilus , for example.
  • the Bifidobacterium bifidum is Bifidobacterium lactis.
  • the composition may be a complete and nutritionally balanced food or pet food. It can also be a dietary supplement, for example. It is preferably addressd to elderly human or elderly pet, or critically ill patients with chronic inflammation.
  • a nutritionally complete pet food can be prepared.
  • the nutritionally complete pet food may be in any suitable form; for example in dried form, semi-moist form or wet form; it may be a chilled or shelf stable pet food product. These pet foods may be produced as is conventional.
  • the prebiotic is provided in the form of plant material, which contains the prebiotic. Suitable plant materials include asparagus, artichokes, onions, wheat, yacon or chicory, or residues of these plant materials.
  • the prebiotic may be provided as an inulin extract or its hydrolysis products commonly known as fructooligosaccharides, galacto-oligosaccarides, xylo-oligosaccharides or oligo derivatives of starch.
  • Extracts from chicory are particularly suitable.
  • the maximum level of prebiotic in the pet food is preferably about 20% by weight; especially about 10% by weight.
  • the prebiotic may comprise about 0.1% to about 5% by weight of the pet food.
  • the chicory may be included to comprise about 0.5% to about 10% by weight of the feed mixture; more preferably about 1% to about 5% by weight.
  • these pet foods may include any one or more of a carbohydrate source, a protein source and lipid source.
  • carbohydrate source is provided in the form of grains, flours and starches.
  • the carbohydrate source may be rice, barley, sorghum, millet, oat, corn meal or wheat flour. Simple sugars such as sucrose, glucose and corn syrups may also be used.
  • the amount of carbohydrate provided by the carbohydrate source may be selected as desired.
  • the pet food may contain up to about 60% by weight of carbohydrate.
  • Suitable protein sources may be selected from any suitable animal or vegetable protein source; for example muscular or skeletal meat, meat and bone meal, poultry meal, fish meal, milk proteins, corn gluten, wheat gluten, soy flour, soy protein concentrates, soy protein isolates, egg proteins, whey, casein, gluten, and the like.
  • the protein source may contain a high quality animal protein.
  • the amount of protein provided by the protein source may be selected as desired.
  • the pet food may contain about 12% to about 70% by weight of protein on a dry basis.
  • the pet food may contain a fat source.
  • Any suitable fat source may be used both animal fats and vegetable fats.
  • the fat source is an animal fat source such as tallow.
  • Vegetable oils such as corn oil, sunflower oil, safflower oil, rape seed oil, soy bean oil, olive oil and other oils rich in monounsaturated and polyunsaturated fatty acids, may also be used.
  • the fat source may include long chain fatty acids.
  • Suitable long chain fatty acids include, gamma linoleic acid, stearidonic acid, arachidonic acid, eicosapentanoic acid, and docosahexanoic acid.
  • Fish oils are a suitable source of eicosapentanoic acids and docosahexanoic acid.
  • Borage oil, blackcurrent seed oil and evening primrose oil are suitable sources of gamma linoleic acid.
  • Rapeseed oil, soybean oil, linseed oil and walnut oil are suitable sources of alpha-linoleic acid.
  • Safflower oils, sunflower oils, corn oils and soybean oils are suitable sources of linoleic acid.
  • Olive oil, rapeseed oil (canola) high oleic sunflower and safflower, peanut oil, rice bran oil are suitable sources of monounsaturated fatty acids.
  • the amount of fat provided by the fat source may be selected as desired.
  • the pet food may contain about 5% to about 40% by weight of fat on a dry basis.
  • the pet food has a relatively reduced amount of fat.
  • the pet food may contain other active agents such as long chain fatty acids.
  • Suitable long chain fatty acids include alpha-linoleic acid, gamma linoleic acid, linoleic acid, eicosapentanoic acid, and docosahexanoic acid.
  • Fish oils are a suitable source of eicosapentanoic acids and docosahexanoic acid.
  • Borage oil, blackcurrent seed oil and evening primrose oil are suitable sources of gamma linoleic acid.
  • Safflower oils, sunflower oils, corn oils and soybean oils are suitable sources of linoleic acid.
  • carbohydrate, protein and lipid sources are not critical and will be selected based upon nutritional needs of the animal, palatability considerations, and the type of product produced. Further, various other ingredients, for example, sugar, salt, spices, seasonings, vitamins, minerals, flavoring agents, gums, and probiotic micro-organisms may also be incorporated into the pet food as desired.
  • a probiotic microorganism may also be added. It may be selected from one or more microorganisms suitable for animal consumption and which is able to improve the microbial balance in the intestine.
  • suitable probiotic micro-organisms include yeast such as Saccharomyces, Debaromyces, Candida, Pichia and Torulopsis , moulds such as Aspergillus, Rhizopus, Mucor , and Penicillim and Torulopsis and bacteria such as the genera Bifidobacterium, Bacteroides, Clostridium, Fusobacterium, Melissococcus, Propionibacterium, Streptococcus, Enterococcus, Lactococcus, Staphylococcus, Peptostrepococcus, Bacillus, Pediococcus, Micrococcus, Leuconostoc, Weissella, Aerococcus, Oenococcus and Lactobacillus .
  • the probiotic micro-organisms may be in powdered, dried form; especially in spore form for micro-organisms which form spores. Further, if desired, the probiotic micro-organism may be encapsulated to further increase the probability of survival; for example in a sugar matrix, fat matrix or polysaccharide matrix. If a probiotic micro-organism is used, the pet food preferably contains about 10 4 to about 10 10 cells of the probiotic micro-organism per gram of the pet food; more preferably about 10 6 to about 10 8 cells of the probiotic micro-organism per gram. The pet food may contain about 0.5% to about 20% by weight of the mixture of the probiotic micro-organism; preferably about 1% to about 6% by weight; for example about 3% to about 6% by weight.
  • the pet food preferably contains proportionally less fat than pet foods for younger pets.
  • the starch sources may include one or more of oat, rice, barley, wheat and corn.
  • a suitable process is extension cooking, although baking and other suitable processes may be used.
  • the dried pet food is usually provided in the form of a kibble.
  • the prebiotic may be admixed with the other ingredients of the dried pet food prior to processing.
  • a suitable process is described in European patent application No 0850569;. If a probiotic micro-organism is used, the organism is best coated onto or filled into the dried pet food.
  • a suitable process is described in European patent application No 0862863.
  • a food composition for human consumption is prepared.
  • This composition may be a nutritional complete formula, a dairy product, a chilled or shelf stable beverage, soup, a dietary supplement, a meal replacement, and a nutritional bar or a confectionery.
  • the nutritional formula may comprise a source of protein.
  • Dietary proteins are preferably used as a source of protein.
  • the dietary proteins may be any suitable dietary protein; for example animal proteins (such as milk proteins, meat proteins and egg proteins); vegetable proteins (such as soy protein, wheat protein, rice protein, and pea protein); mixtures of free amino acids; or combinations thereof. Milk proteins such as casein, whey proteins and soy proteins are particularly preferred.
  • the composition may also contain a source of carbohydrates and a source of fat.
  • the fat source preferably provides about 5% to about 55% of the energy of the nutritional formula; for example about 20% to about 50% of the energy.
  • the lipids making up the fat source may be any suitable fat or fat mixtures. Vegetable fats are particularly suitable; for example soy oil, palm oil, coconut oil, safflower oil, sunflower oil, corn oil, canola oil, lecithins, and the like. Animal fats such as milk fats may also be added if desired.
  • a source of carbohydrate may be added to the nutritional formula. It preferably provides about 40% to about 80% of the energy of the nutritional composition. Any suitable carbohydrates may be used, for example sucrose, lactose, glucose, fructose, corn syrup solids, and maltodextrins, and mixtures thereof. Dietary fibre may also be added if desired. If used, it preferably comprises up to about 5% of the energy of the nutritional formula.
  • the dietary fibre may be from any suitable origin, including for example soy, pea, oat, pectin, guar gum, gum arabic, and fructooligosaccharides. Suitable vitamins and minerals may be included in the nutritional formula in an amount to meet the appropriate guidelines.
  • One or more food grade emulsifiers may be incorporated into the nutritional formula if desired; for example diacetyl tartaric acid esters of mono- and di-glycerides, lecithin and mono- and di-glycerides. Similarly suitable salts and stabilisers may be included.
  • the nutritional formula is preferably enterally administrable; for example in the form of a powder, tablet, capsule, a liquid concentrate, solid product or a ready-to-drink beverage. If it is desired to produce a powdered nutritional formula, the homogenised mixture is transferred to a suitable drying apparatus such as a spray drier or freeze drier and converted to powder.
  • a suitable drying apparatus such as a spray drier or freeze drier and converted to powder.
  • a nutritional composition comprises a milk based cereal together with a prebiotic formulation.
  • the milk based cereal is an infant cereal which acts as a carrier for the prebiotic formulation.
  • a usual food product may be enriched with at least one prebiotic according to the present invention.
  • the amount of the composition added is preferably at least about 0.01% by weight.
  • FIG. 1 Effect of feeding 8 g of short-chain fructooligosaccharides (FOS) on viable counts of bifidobacteria in fresh faecal samples of nursing home elderly. Time points were before (2), during (3), and after (4) period of 3 weeks of FOS intake.
  • A Individual values.
  • B Box plots. Boxes indicate 25 th and 75 th percentiles, solid lines inside box indicate median values, and t-shapes 5 th and 95 th percentiles; solid dot indicates outlying value.
  • FIG. 2 Effect of fructooligosaccharides (FOS) on bifidobacteria in adults and elderly.
  • Reference 24 Bohnik Y et al. J Nutr 1999; 129:113
  • Reference 25 Merenne E et al. J Nutr 2000;130:1197) response to 8 g FOS/day in 20-50 year old adults
  • Reference 26 Garson G R J Nutr 1999;129:1438S
  • Reference 27 Korean H P et al.
  • FIG. 3 Differences in IL-6 mRNA expression in peripheral blood mononuclear cells (PBMC) of elderly supplemented with FOS.
  • IL-6 mRNA expression in PBMC was measured by reverse transcription-polymerase chain reaction (RT-PCR). Estimation of quantitative changes were done by scanning band intensity using NIH-image program and calculated as density ratio of IL-6 mRNA to ⁇ -actin mRNA in percent. Time points were before (2), during (3), and after (4) period of 3 weeks of FOS intake.
  • A Representative image of individual changes from ethidium bromide stained gels.
  • the study was a pretest/posttest study of 19 elderly nursing home patients (see study scheme).
  • MNA Mini Nutritional Assessemnt
  • FOS short chain fructooligosaccharides
  • Stool samples were collected at day 0, 21, 42 and 63 from each subject.
  • the stool samples were immediately (within 30 minutes) placed in an anaerobic jar and kept at 4° C. until analysis (a maximum of 6 hours).
  • Hundred fold serial dilutions were performed in pre-reduced Ringer solution containing 0.5% of cystein, from ⁇ 2 to ⁇ 8.
  • Petri dishes of various media were inoculated and incubated for 48 hr at 37° C. in anaerobic atmosphere using Anaerocult A (Merck, Darmstadt, Germany), except for Enterococci and Enterobacteriacea incubated for 24 hr at 37° C. in aerobic atmosphere.
  • Bacteria were detected on selective or half-selective media as follows: Enterobacteriacea on Drigalski medium (Sanofi Diagnostics Pasteur, France), Bifidobacteria on Eugon Tomato medium (Wadsworth Anaerobic Bacteriology Manual, V. Suter, D. Citron and S. Finegold Third edition), Lactobacilli in MRS (Difco, MI.
  • antibiotics Phosphomycine (79.5 mg/l)+Sulfamethoxazole (0.93 mg/l)+Trimethoprime (5 mg/l)
  • Clostridium perfringens on NN agar Liowbury and Lilly, 1995
  • Bactero ⁇ des on Schaedler Neo-Vanco medium BioMérieux, Marcy-I'Etoile, France
  • Enterococci on Azide agar Difco).
  • Lactobacilli and Bifidobacteria strains were identified by microscopy, and biochemically using the API gallery system (BioMérieux), API 50 CHL gallery for lactobacilli, and API ID 32A gallery for Bifidobacteria respectively. Bacterial counts are expressed as log 10 colony-forming units (CFUs) per gram of fresh faecal sample, with a detection limit at 3.30 cfu/g.
  • Faecal pH was measured just after emission by the nurse at three different points in crude faeces with a microelectrode (Orion). The mean pH was calculated for each time point.
  • Serum albumin, transthyretin (prealbumin), C-reactive protein, and a, acid glycoprotein concentrations were analyzed by immuno-nephelometry with a Behring Nephelometer (methods and reagents from Behring, Marburg, Germany).
  • Serum folate and vitamin B 12 (cobalamin) concentrations were analyzed by radioimmunoassay (Dual Count, Diagnostic Product Corporation, Los Angeles, Calif., USA).
  • Serum albumin normal range 35-55 g/L
  • transthyretin prealbumin; normal range 0.16-0.40
  • the acute phase proteins ⁇ 1-acid glycoprotein (normal range 0.5-1.3)
  • C-reactive protein normal values ⁇ 10 mg/L and abnormal values for elderly >20 mg/L
  • 2 men elevated C-reactive protein levels
  • 2 women low albumin levels with normal transthyretin and borderline C-reactive protein
  • Bacterial counts of bifidobacteria were increased by a mean of 2.8 ⁇ 0.57 log 10 CFU during the 3 weeks of FOS supplementation (p ⁇ 0.001), but Bactero ⁇ des counts were also increased p ⁇ 0.032) (see tables 2a & 2b and FIG. 1 ).
  • Phagocytic activity of granulocytes and monocytes were significantly decreased by the ingestion of FOS: Phagocytic activity expressed as median fluorescent intensity changed for granulocytes from 130 ⁇ 10 to 52 ⁇ 2 (p ⁇ 0.001) and for monocytes from 75 ⁇ 5 to 26 ⁇ 2 (p ⁇ 0.001).
  • Serum vitamin B 12 were at 271 ⁇ 143 ng/L before supplementation and at 289 ⁇ 160 ng/L during supplementation. Three subjects, however, were deficient in vitamin B 12 . Two subjects returned to normal status during the study while the other remained deficient throughout study. Serum folate levels were at 5.9 ⁇ 2.0 ug/L before supplementation and at 5.7 ⁇ 1.9 ug/L during supplementation.
  • the present study confirms the positive effect of FOS supplementation on bifidobacteria observed in adults and elderly ( FIG. 2 ), indicating that elderly respond to prebiotic (FOS) intake by an increase in bifidobacteria like younger adults or even better if the bifidobacteria counts are low. Further 8 g of FOS or less seems to be sufficient to achieve a maximal effect on bifidobacteria counts. While from different studies a dose response seems to be present, single studies indicate that above a threshold of 4-5 g/day a maximal response is obtained and the increase in bifidobacteria seems to be more dependent of the initial number ( FIGS. 1 and 2 ).
  • a food supplement was prepared by mixing or blending fructooligosaccharide with inulin in the proportions by weight of about 70% fructooligosaccharide to about 30% inulin.
  • the resulting prebiotic mixture may be added or blended with any suitable carrier, for example a fermented milk, a yogurt, a fresh cheese, a renneted milk, a confectionery bar, breakfast cereal flakes or bars, a drink, milk powder, soy-based product, non-milk fermented product or a nutritional supplement for clinical nutrition.
  • a feed mixture is made up of about 58% by weight of corn, about 5.5% by weight of corn gluten, about 22% by weight of chicken meal, 2,5% dried chicory, 1% carnitine, salts, vitamins and minerals making up the remainder.
  • the fed mixture is fed into a preconditioner and moistened.
  • the moistened feed is then fed into an extruder-cooker and gelatinised.
  • the gelatinised matrix leaving the extruder is forced through a die and extruded.
  • the extrudate is cut into pieces suitable for feeding to dogs, dried at about 110° C. for about 20 minutes, and cooled to form pellets.
  • This dry dog food is particularly intended for decreasing inflammatory process and/or abnormal activation of non specific immune parameters, such as phagocytes.
  • a mixture is prepared from 56% of poultry carcass, pig lungs and pig liver (ground), 13% of fish, 16% of wheat flour, 2% of plasma, 10.8% of water, 2.2% of dyes, 1% of semi refined kappa carrageenan, inorganic salts and 9% oil rich in monounsaturated fatty acids (olive oil) and 3% chicory.
  • This mixture is emulsified at 12° C. and extruded in the form of a pudding which is then cooked at a temperature of 90° C. It is cooled to 30° C. and cut in chunks.

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US9427000B2 (en) 2005-05-31 2016-08-30 Mars, Incorporated Feline probiotic lactobacilli composition and methods
US9580680B2 (en) 2003-12-19 2017-02-28 Mars, Incorporated Canine probiotic bifidobacterium pseudolongum
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US10681922B2 (en) 2003-12-01 2020-06-16 Mars, Incorporated Methods of enhancing the gastrointestinal health of a companion animal
US9580680B2 (en) 2003-12-19 2017-02-28 Mars, Incorporated Canine probiotic bifidobacterium pseudolongum
US9821015B2 (en) 2003-12-19 2017-11-21 Mars, Incorporated Methods of use of probiotic bifidobacteria for companion animals
US9415083B2 (en) 2004-05-10 2016-08-16 Mars, Incorporated Method for decreasing inflammation and stress in a mammal
US9404162B2 (en) 2005-05-31 2016-08-02 Mars, Incorporated Feline probiotic bifidobacteria and methods
US9427000B2 (en) 2005-05-31 2016-08-30 Mars, Incorporated Feline probiotic lactobacilli composition and methods
US10709156B2 (en) 2008-07-07 2020-07-14 Mars, Incorporated Pet supplement and methods of making
US10104903B2 (en) 2009-07-31 2018-10-23 Mars, Incorporated Animal food and its appearance
US11122821B2 (en) 2016-12-15 2021-09-21 Societe Des Produits Nestle S.A. Compositions and methods that modulate bacteria in a companion animal

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EP1383514B1 (en) 2007-01-03
DE60217319T3 (de) 2019-04-18
ES2278021T3 (es) 2007-08-01
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ATE350045T1 (de) 2007-01-15
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AR033439A1 (es) 2003-12-17
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CA2442053C (en) 2012-07-10

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